Detection of pork adulteration in beef with ladder-shape melting temperature isothermal amplification (LMTIA) assay

ABSTRACT The ladder-shape melting temperature isothermal amplification (LMTIA) is a newly developed technology, and the objective of this study is to establish the LMTIA assay for detection of pork in beef. The LMTIA primers were designed with the prolactin receptor gene of Sus scrofa as the target. The LMTIA reaction system was optimized, and the performance of the LMTIA assay in specificity, sensitivity and detection limit (LOD) was determined. Our results showed that the LMTIA assay was able to specifically detect 10 ng genomic DNA (gDNAs) of Sus scrofa, the sensitivity of the LMTIA assay was 1 ng genomic DNA of Sus scrofa, and the LMTIA assay was able to detect the pork adulterated in beef with 0.1% LOD, which was the same as that of reported PCR assay or LAMP assay. This study holds a promise for facilitating the surveillance of the commercial adulteration of pork in beef.